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Mishra, K. ; Fuenzalida Werner, J.P. ; Pennacchietti, F. [extern]* ; Janowski, R. ; Chmyrov, A. ; Huang, Y. ; Zakian Dominguez, C.M. ; Klemm, U. ; Testa, I. [extern]* ; Niessing, D. ; Ntziachristos, V. ; Stiel, A.-C.

Genetically encoded photo-switchable molecular sensors for optoacoustic and super-resolution imaging.

Nat. Biotechnol., DOI: 10.1038/s41587-021-01100-5 (2021)
Publ. Version/Full Text Research data DOI PMC
Open Access Gold (Paid Option)
Creative Commons Lizenzvertrag
Reversibly photo-switchable proteins are essential for many super-resolution fluorescence microscopic and optoacoustic imaging methods. However, they have yet to be used as sensors that measure the distribution of specific analytes at the nanoscale or in the tissues of live animals. Here we constructed the prototype of a photo-switchable Ca2+ sensor based on GCaMP5G that can be switched with 405/488-nm light and describe its molecular mechanisms at the structural level, including the importance of the interaction of the core barrel structure of the fluorescent protein with the Ca2+ receptor moiety. We demonstrate super-resolution imaging of Ca2+ concentration in cultured cells and optoacoustic Ca2+ imaging in implanted tumor cells in mice under controlled Ca2+ conditions. Finally, we show the generalizability of the concept by constructing examples of photo-switching maltose and dopamine sensors based on periplasmatic binding protein and G-protein-coupled receptor-based sensors.Calcium and other analytes can be imaged at super-resolution and in vivo with photo-switchable sensors.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Fluorescent Proteins; Calcium Indicator; Mitochondria; Microdomains; Microscopy; Mesoscopy; Mechanism
ISSN (print) / ISBN 1087-0156
e-ISSN 1546-1696
Journal Nature Biotechnology
Publisher Nature Publishing Group
Publishing Place New York, NY
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Biological and Medical Imaging (IBMI)
Institute of Structural Biology (STB)
Grants VR starting grant
ERC
China Scholarship Council
European Union Horizon 2020 research and innovation program
DFG